Opiate Modulates Lymphocyte Trafficking into the CNS in TB Meningitis

描述（由申请人提供）：在全球范围内，结核病（TB）仍然是一个主要的公共卫生危机，艾滋病毒的流行大大加剧了这一危机。临床上，结核分枝杆菌是艾滋病患者最重要的条件致病菌，而中枢神经系统（CNS）感染是结核病最具破坏性的并发症。与HIV感染一样，药物滥用被认为是M.结核虽然已经有许多关于阿片类药物对其他细胞内微生物的发病机制的影响的研究，但关于阿片类药物对淋巴细胞运输的影响知之甚少，关于阿片类药物对CNS防御M的影响也一无所知。结核在中枢神经系统内，M.结核病引起神经炎症反应，但参与这种反应的细胞和介质在很大程度上是不确定的。虽然致敏T淋巴细胞在防御许多CNS感染中起关键作用，但T淋巴细胞响应感染因子（包括M.结核病，尚未明确。最近，一种新的成像系统，利用转基因小鼠表达荧光素酶的开发，使研究人员能够定量跟踪“体内”的运输模式的T淋巴细胞进入中枢神经系统的多发性硬化症的动物模型。在本申请中提出的研究的总体目标是适应这种创新的方法，以评估阿片类药物依赖性的影响T淋巴细胞贩运到中枢神经系统在脑内的挑战与结核杆菌。这种定量成像系统的使用将使我们能够测试的中心假设，阿片类药物依赖损害贩运的卡介苗（BCG）致敏的T淋巴细胞进入中枢神经系统，从而增加中枢神经系统结核病的严重程度。为了验证这一假设，设计了解决以下两个具体目标的实验：具体目标1：采用新的定量成像技术来确定BCG致敏的T淋巴细胞是否响应于分枝杆菌感染而进入CNS。据推测，卡介苗接种将通过卡介苗致敏的T淋巴细胞在接种结核杆菌后迅速浸润到中枢神经系统来保护小鼠免受中枢神经系统结核病的侵害。具体目标2：确定吗啡给药是否增强分枝杆菌感染的神经发病机制，并损害T淋巴细胞运输到中枢神经系统。利用上述CNS TB模式和T细胞成像技术，将阐明吗啡给药对神经发病机制的影响。据推测，吗啡依赖将加速死亡率和增加CNS损伤，由于感染和过继转移BCG致敏的T淋巴细胞的运输将在吗啡依赖小鼠中显着受损。

DESCRIPTION (provided by applicant): Globally, tuberculosis (TB) remains a major public health crisis, which has been dramatically fueled by the HIV epidemic. Clinically, Mycobacterium tuberculosis is the most important opportunistic pathogen in AIDS patients, and infection of the central nervous system (CNS) is the most devastating complication of TB. As in the case of HIV infection, drug abuse has been regarded as a potential cofactor in the pathogenesis of M. tuberculosis. While there have been numerous studies on the effects of opiates on the pathogenesis of other intracellular microbes, little is known about the influence of opiates on lymphocyte trafficking and nothing is known about the impact of opiates on defense of the CNS against M. tuberculosis. Within the CNS, M. tuberculosis elicits a neuroinflammatory response, but the cells and mediators involved in this response are largely undefined. Although sensitized T lymphocytes play a critical role in defense against many CNS infections, the process of T lymphocyte entry into the brain in response to infectious agents, including M. tuberculosis, has not been delineated. Recently, a novel imaging system utilizing transgenic mice expressing luciferase was developed allowing researchers to track quantitatively "in vivo" the trafficking pattern of T lymphocytes into the CNS in an animal model of multiple sclerosis. The overall goal of the research proposed in this application is to adapt this innovative methodology to assess the effects of opiate dependence on T lymphocyte trafficking into the CNS in response to intracerebral challenge with tubercle bacilli. The use of this quantitative imaging system will allow us to test the central hypothesis that opiate dependence impairs the trafficking of Bacille-Calmette-Gu¿rin (BCG)-sensitized T lymphocytes into the CNS thereby increasing the severity of CNS TB. To test this hypothesis, experiments have been designed which address the two specific aims outlined below: Specific aim 1: Adapt a novel quantitative imaging technique to determine whether BCG- sensitized T lymphocytes traffic into the CNS in response to mycobacterial infection. It is hypothesized that BCG vaccination will protect mice against CNS TB through a rapid infiltration of BCG- sensitized T lymphocytes into the CNS following ic inoculation of tubercle bacilli. Specific aim 2: Determine whether morphine administration enhances the neuropathogenesis of mycobacterial infection and impairs T lymphocyte trafficking into the CNS. Using the CNS TB paradigm above and T cell imagining technology, the effects of morphine administration on the neuropathogenesis will be elucidated. It is hypothesized that morphine dependence will accelerate mortality and increase CNS damage due to infection and that the trafficking of adoptive transferred BCG- sensitized T lymphocytes will be markedly impaired in morphine-dependent mice.